Method for preparing ginkgolide B by using Tremella fungus thalline as catalyst

A technology of ginkgolide and tremella fungus, applied in the field of bioengineering, can solve the problems of producing ginkgolide and difficulty in mass production

Active Publication Date: 2016-07-06
BEIJING LYUKE TIANCHENG BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

And there are only nine articles about the preparation of ginkgolide B (a new method for extracting ginkgolide B from ginkgo biloba or ginkgo leaf extract, application number 200510063407.X; a kind of separation of ginkgolide from ginkgolide mixture The method of B, application number: CN201010608847.X, etc.), the ginkgolides contain ginkgolides A, B, C, M and other homologues in the ginkgolides, these patents adopt separation technology to separate ginkgolides from the ginkgolide mixture B, it is difficult to produce ginkgolides in large quantities in industry B

Method used

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  • Method for preparing ginkgolide B by using Tremella fungus thalline as catalyst

Examples

Experimental program
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Effect test

Embodiment 1

[0014] The preparation of embodiment 1 test tube bacterial classification

[0015] Preparation of Potato Dextrose Agar Medium: Weigh 200g of potatoes, cut them into slices, add 1000mL of water, boil for 30 minutes, filter, dilute the filtrate to 1000mL, add 20g of agar, heat until the agar is completely melted, and pack into 18×200mm 15mL medium per tube, sterilized at 120°C for 30 minutes, cooled to 50°C, placed on a slope, the length of the slope is half the length of the test tube; cut the preserved Tremella fungus isolated from the fruiting body of Tremella into 4× 4mm small block bacteria, pick a small piece and transfer it to a test tube containing potato dextrose agar medium, culture at 20°C for 20 days, and prepare the test tube slant strain, and store the test tube slant at 4°C for later use:

Embodiment 2

[0016] Embodiment 2 contains the preparation of the fermented liquid of ginkgolide B

[0017] 1. Preparation of strains in liquid shake flasks

[0018] The formula of liquid shake flask culture medium is (unit is g / L) glucose 5, corn flour 5, peptone 1, KH 2 PO 4 1. MgSO 4 1. The initial pH value is 5.0. A total of 5500mL was prepared, divided into 250mL Erlenmeyer flasks, each bottle was 60mL, a total of 84 bottles were sterilized at 100°C for 60 minutes. A total of 5500mL was prepared, divided into 250mL triangular flasks, each bottle was 60mL, a total of 84 bottles, the preserved Tremella test tube slant strain isolated from the fruiting body of Tremella yunnanensis was cut into small pieces of 4×4mm size, and 3 pieces were picked for transfer Put it into a 250mL Erlenmeyer flask filled with shaker culture medium, and place it on a shaker at 22°C and 90 rpm for 24 hours.

[0019] 2. Preparation of seed tank strains

[0020] The 75L seed tank is filled with 45L seed cu...

Embodiment 3

[0025] Embodiment 3 contains the preparation of the fermented liquid of ginkgolide B

[0026] 1. Production of strains in shake flasks

[0027] The formula of the shake flask medium is (in g / L): glucose 20, corn flour 15, peptone 6, KH 2 PO 4 3. MgSO 4 2. The initial pH value is 6.5, a total of 4500mL is prepared, divided into 250mL Erlenmeyer flasks (total 50 bottles), each bottle is 90mL, and sterilized at 120°C for 40 minutes. After cooling, cut the preserved tremella test tube slant strains separated from Shanxi tremella fruiting bodies into small pieces of 4×4mm size, and cut the test tube slant strains into small pieces of 4×4mm size, pick 4 pieces and transfer them into Place the Erlenmeyer flask with shaker culture medium at 25°C on a shaker at 120 rpm for 36 hours.

[0028] 2. Preparation of seed tank strains

[0029] 100L seed tank is filled with 50L seed culture medium, wherein the formula of the seed tank culture medium is (unit is g / L): glucose 20, corn flour...

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Abstract

The invention discloses a method for synthesizing ginkgolide B by using tremella (Stereum hirsutum) thalline as a catalyst to transform a mixture of ginkgolides, and belongs to the field of biotechnology. The present invention expands and cultivates Tremella fungus strains, separates them from test tube slopes, liquid shake flasks, seed culture, and thallus, uses the thallus as a catalyst, converts ginkgolides to synthesize ginkgolide B, and the transformation solution is centrifuged, ethyl acetate Extraction and absolute alcohol crystallization separate ginkgolide B from the fermentation broth. The purity of the ginkgolide B obtained by the method is greater than or equal to 99%, and the conversion rate of the ginkgolide is greater than or equal to 90%.

Description

technical field [0001] The invention relates to the technical field of bioengineering, and relates to a new process for preparing ginkgolide B by transforming ginkgo biloba extracts through liquid culture of Tremella fungus strains. Background technique [0002] Studies have proved that ginkgolide is one of the main active ingredients in Ginkgo biloba, and is a specific and effective platelet activating factor (Platelet activating factor PAF) receptor antagonist. Platelet activating factor is an endogenous phospholipid secreted by platelets and various inflammatory tissues. It is the most effective platelet aggregation inducer found so far, and is closely related to the occurrence and development of many diseases. Ginkgolides, as specific antagonists of PAF receptors, have a wide range of pharmacological effects. Studies have found that ginkgolide B has protective effects on the central nervous system and ischemic injury, anti-shock, anti-allergic, antibacterial and anti-in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/18C12R1/645
Inventor 张志才
Owner BEIJING LYUKE TIANCHENG BIO TECH
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